Number Of Follicles Research Articles

P-590 Should the ovarian ultrasound markers for the diagnosis of PCOS be revised?

Abstract Study question Should the cut-off values for follicle number and ovarian volume in PCOS be revised, and should they be adjusted based on age or BMI? Summary answer The cut-off for follicle number per ovary can be maintained (age specific, not BMI adjusted), while for ovarian volume, it should be lowered or omitted. What is known already The 2023 International Guideline for PCOS concluded that the quality of evidence for recommendations regarding the ultrasound diagnosis of PCOS (FNPO ≥ 20 and/or ovarian volume (OV) ≥10 mL, in one or both ovaries) is low. It advises to regularly revise thresholds with advancing ultrasound technology and to investigate the impact of age on diagnostic accuracy. Consideration of other biological factors such as BMI is also warranted. Study design, size, duration Retrospective cross-sectional study, including data from 2001-2023, obtained through standardized endocrinological screening at the Department of Reproductive Endocrinology in the Erasmus Medical Center (Rotterdam, The Netherlands). Women diagnosed with PCOS during the screening were eligible for inclusion in the PCOS group. Women who underwent the same screening, had regular menstrual cycles, exhibited normal hormonal values, and showed no endocrinological pathology were eligible for inclusion as controls. Participants/materials, setting, methods A total of 2492 women (15-50 years) with PCOS and 152 controls were included. PCOS was diagnosed according to the 2003 Rotterdam criteria and the 2018 International Guideline. Exclusion criteria included: age < 15 or > 50 years, presence of any cysts (>10 mm) in one/both ovaries, use of oral contraceptives, and missing ultrasound data. The ovarian ultrasound markers were stratified by age in 5-year intervals. Receiver Operating Characteristic (ROC) analyses were performed for optimal cut-offs. Main results and the role of chance In our cohort, most women with PCOS up to age 35 exhibit a TFC ≥ 20, followed by a decline in FNPO above the age of 35. In a subgroup of women aged 35-40 (n = 24 PCOS, n = 24 controls, matched by age, assessed with ultrasound > 8 MHz), we found optimal cut-offs of 14.5 follicles (Left FNPO) and 14 follicles (Right FNPO), with AUC values of 0.94 and 0.93. Median OV in every age group was below the 10 mL cut-off. In women aged 20-35 (n = 75 PCOS, 75 controls, ultrasound > 8 MHz), optimal cut-offs were 6.5 mL (Left OV) and 7.4 mL (Right OV), with AUC values of 0.70 and 0.78. For 35-40 years, cut-offs were 8.0 mL (Left OV) and 7.2 mL (Right OV), including 24 women PCOS and 24 age-matched controls, with AUC values of 0.68 and 0.62. In our cohort 19/2413 women with PCOS (0.8%) would not have received the diagnosis PCOS if OV was not considered as a marker for PCOM. No clinically significant correlation between BMI and FNPO or OV was observed. Limitations, reasons for caution The optimal cut-off values should be interpreted with caution due to the small sample sizes in the ROC analyses. The control group includes women with isolated PCOM. There is an ongoing debate about whether they should be excluded in such analyses. Wider implications of the findings We suggest lowering or omitting the OV cut-off for PCOM detection and establishing age-specific FNPO cut-offs. Future research on ovarian markers in PCOS, should prioritize women with isolated ovulatory dysfunction or hyperandrogenism, aligning with the 2023 International PCOS Guideline recommendation to exclusively investigate PCOM in this subgroup. Trial registration number not applicable

The Effect of Progesterone Elevation on the Day of Trigger Administration: A Review of the Literature.

Since the advent of assisted reproductive technology, different variables have been shown to affect pregnancy outcomes. One of the most prevalent studied events is the premature rise in serum progesterone concentrations on the day of trigger administration during cycles of ovarian stimulation. This phenomenon, classically known as premature luteinization, has been observed significantly for decades and has been linked to adverse pregnancy outcomes and lower live birth rates. Ultimately, a quest to find a precise serum progesterone concentration cut-off value that can be effectively used to predict pregnancy outcomes prior to trigger administration is still underway. The purpose of the current research was to study the available literature on the relationship between serum progesterone on the day of trigger administration in controlled ovarian stimulation cycles used for IVF in an attempt to identify a cut-off serum progesterone concentration that can be used to effectively predict future pregnancy outcomes in fresh transfers. This study is a review of the literature and is based on information and data gathered from 36 published articles. The majority of the literature shows that a serum progesterone concentration cut-off of 1.5 ng/ml (4.77 nmol/L) can be used prior to trigger administration to effectively predict pregnancy outcomes. Premature progesterone elevation on the day or prior to the trigger administration is associated with adverse pregnancy outcomes in IVF cycles. Other factors such as follicle number, serum concentration of other hormones, and ovarian response to ovarian stimulation should also be considered to predict the success of IVF protocols.

Ovarian hyperplasia linked to a mutation in MAN1A2 in a cow with excessive follicular growth and functional oocytes.

Here we report the case of a cow with two ovaries that each exhibited hyperplasia but that otherwise had normal gross morphology. Both ovaries had a large number of tertiary follicles on the ovarian surface. Oocytes from one ovary were studied in more detail. The transcriptome was largely similar to other oocytes. Oocytes could undergo cleavage at a rate consistent with other oocytes and result in blastocyst-stage embryo formation after in vitro maturation and fertilization. Review of the literature from cattle and other species did not reveal reports of a similar type of spontaneous ovarian abnormality. Whole genome sequencing revealed many single nucleotide polymorphisms with predicted large effects on protein structure that could potentially be causative for the phenotype. The variant considered most likely to cause the observed alteration in ovarian function was a mutation in the glycoprotein-modifying enzyme MAN1A2.

Chemotherapy drug combinations induced maternal ovarian damage and long-term effect on fetal reproductive system in mice

With the postponement of female reproductive age and the higher incidence of cancer in young people, fertility preservation has become increasingly important in childbearing age. Chemotherapy during pregnancy is crucial for maternal cancer treatments and fetal outcomes. It is a need to further study ovarian damage caused by chemotherapy drug combinations and long-term effects on offspring development, and a detailed understanding of side effects of chemotherapy drugs.In this study, chemotherapy drug combinations significantly impacted on ovarian function, especially epirubicin/cyclophosphamide (EC) combination led to an unbalance in the development of the left and right ovary. Exposure to EC and cisplatin/paclitaxel (TP) increased the number of progenitor follicles while decreased the count of antral follicles and corpora luteum. As to the estrus cycle, EC exposure resulted in a longer estrus period and diestrus period, while TP exposure only extended the diestrus period. EC and TP affected steroid biosynthesis by reducing the expression of SF1 and P450arom.γ-H2AX was detected in both EC and TP exposure groups.As to the impact on the offspring from 4T1 tumor-bearing pregnant mice injected with EC, no significant difference was observed in the physical and neurological development compared to the control, but the ovarian weights, estrus cycles of the offspring were significantly different. Chemotherapy drug combinations exhibit ovarian toxicity, not only causing direct damage on the follicle cells but also disrupting steroid biosynthesis. The reproductive system of offspring from maternal tumor-bearing mice exposed to chemotherapy drugs was observed disorder, but the concrete mechanism still needs further exploration.

Discovery and evaluation of novel spiroheterocyclic protective agents via a SIRT1 upregulation mechanism in cisplatin-induced premature ovarian failure

Currently, no effective treatment exists for premature ovarian failure (POF). To obtain compounds with protective effects against POF, we aimed to design and synthesize a series of spiroheterocyclic protective agents with a focus on minimizing toxicity while enhancing their protective effect against cisplatin-induced POF. This was achieved through systematic modifications of Michael receptors and linkers within the molecular structure of 1,5-diphenylpenta-1,4-dien-3-one analogs. To assess the cytotoxicity and activity of these compounds, we constructed quantitative conformational relationship models using an artificial intelligence random forest algorithm, resulting in R2 values exceeding 0.87. Among these compounds, j2 exhibited optimal protective activity. It significantly increased the survival of cisplatin-injured ovarian granulosa KGN cells, improved post-injury cell morphology, reduced apoptosis, and enhanced cellular estradiol (E2) levels. Subsequent investigations revealed that j2 may exert its protective effect via a novel mechanism involving the activation of the SIRT1/AKT signal pathway. Furthermore, in cisplatin-injured POF in rats, j2 was effective in increasing body, ovarian, and uterine weights, elevating the number of follicles at all levels in the ovary, improving ovarian and uterine structures, and increasing serum E2 levels in rats with cisplatin-injured POF. In conclusion, this study introduces a promising compound j2 and a novel target SIRT1 with substantial protective activity against cisplatin-induced POF.

Superovulating cattle with corifollitropin-alpha, a long-acting recombinant human FSH (rhFSH): Dose-response, half-life, effects on the ovaries, and embryo outcomes

The aim of this study was to evaluate the superestimulatory and superovulatory responses of cattle treated with corifollitropin-alpha, a long-acting human recombinant FSH (rhFSH). In the first and second experiments, we used Nelore (Bos indicus) heifers previously submitted to follicular wave suppression by active immunization against GnRH. In Experiment 1 (a dose-response study), heifers (n = 20) were randomly allocated into five groups, which received placebo (saline) or a single sc dose of 7.5, 15.0, 22.5 or 30.0 μg rhFSH. The heifers were subjected to daily ovarian scan and blood sampling during 11 days. We observed group, time, and group x time effects (P<0.0001) for both average follicle size and circulating FSH concentrations, with a strong correlation (R = 0.82, P<0.0001) between the area under curve (AUC) for both parameters. The peak concentration of FSH 24h after treatment and average follicle size at all timepoints, however, were similar (P>0.05) between groups 22.5 and 30.0 μg. In Experiment 2, heifers (n = 18) were allocated into three groups, which received (0h) either placebo (control), 25 μg rhFSH or 130 mg pFSH (Folltropin). There was no difference (P>0.05) in average follicle size at any moment, as well as in intrafollicular E2 at 120h or in plasma P4 seven days later between groups rhFSH and pFSH. In Experiment 3, cycling Nelore heifers (n = 20) were subjected to a wave synchronization protocol and superovulated (day 0) using a standard pFSH protocol (120 mg split in eight decreasing im doses) or with a single sc injection of 20 μg rhFSH. The number of follicles >7 mm on day 4 did not differ (P=0.4370). Heifers receiving rhFSH had greater average follicle size on day 4 (P=0.0005), ovulation rate (P<0.0001), and number of CL (P=0.0155), as well as a trend towards a greater number of ova (P=0.07) and viable embryos (P=0.0590). In Experiment 4, superovulation was induced with a single sc injection of 25 μg rhFSH in Girolando and Nelore cows and heifers (n = 20). None of the embryo yield endpoints differed between the two breeds (P>0.05). In conclusion, cattle superstimulation and superovulation can be successfully induced with a single dose of a long-acting rhFSH (corifollitropin-alpha).

Macroscopic morphological analysis of different stages of Cyclic Corpus Luteum in Indian buffalo (Bubalus bubalis)

The current study was undertaken with the aim to characterize the macroscopic morphological and morphometrical features of cyclic corpus luteum (CL; n=40) and CL of pregnancy (n=10) in buffalo. The four stages of cyclic CL were interpreted after ovarian analysis i.e., c olor, consistency, vasculature of CL, number and size of follicles into early (Stage I, 1 to 5 days, n=10), mid (Stage II, 6 to 11 days, n=10), late luteal phase (Stage III, 12 to 16 days, n=10) and follicular phase (Stage IV, 17 to 20 days, n=10). In Stage I, it was slightly protruded from the surface of ovary, bloody in appearance due to increased blood congestion, soft in consistency and termed as corpus haemorrhagicum. In Stage II, initially CL was bright red in color, later fleshy in color and soft in consistency. In Stage III, it was shrunken to great extent and pale yellow to creamish in color due to reduced vascularity. At Stage IV it was shrunken and rigid; texture became firmer, completely condensed into small whitish in color due to complete loss of vascularity. It varied in size and weight as well during the varying stages of estrus cycle depicting changes in its morphology. Therefore, by recording the macroscopic observations on cyclic CL and CL of pregnancy, it was further characterized into different stages.

Slit1 inhibits ovarian follicle development and female fertility in mice†.

Previous in vitro studies have suggested that SLIT ligands could play roles in regulating ovarian granulosa cell proliferation and gene expression, as well as luteolysis. However, no in vivo study of Slit gene function has been conducted to date. Here, we investigated the potential role of Slit1 in ovarian biology using a Slit1-null mouse model. Female Slit1-null mice were found to produce larger litters than their wild-type counterparts due to increased ovulation rates. Increased ovarian weights in Slit1-null animals were found to be due to the presence of greater numbers of healthy antral follicles with similar numbers of atretic ones, suggesting both an increased rate of follicle recruitment and a decreased rate of atresia. Consistent with this, treatment of cultured granulosa cells with exogenous SLIT1 induced apoptosis in presence or absence of follicle-stimulating hormone, but had no effect on cell proliferation. Although few alterations in the messenger RNA levels of follicle-stimulating hormone-responsive genes were noted in granulosa cells of Slit1-null mice, luteinizing hormone target gene mRNA levels were greatly increased. Finally, increased phospho-AKT levels were found in granulosa cells isolated from Slit1-null mice, and SLIT1 pretreatment of cultured granulosa cells inhibited the ability of both follicle-stimulating hormone and luteinizing hormone to increase AKT phosphorylation, suggesting a mechanism whereby SLIT1 could antagonize gonadotropin signaling. These findings therefore represent the first evidence for a physiological role of a SLIT ligand in the ovary, and define Slit1 as a novel autocrine/paracrine regulator of follicle development.

Role of DNA damage response in cyclophosphamide-induced premature ovarian failure in mice.

To investigate the DNA damage response (DDR) in a cyclophosphamide (CTX)-induced mouse model of premature ovarian failure (POF). The POF model was established by injecting mice with CTX. The body, ovarian weights, the estrus cycle, and pathological changes of the ovaries were recorded. The serum levels of 17 β-estradiol (E2) and follicle-stimulating hormone (FSH) were measured. The expression of Ki67, β-galactosidase (β-gal), p21, p53, γH2AX, and pATM in ovarian tissues was detected by immunohistochemistry. The expression of β-gal, γH2AX, and pATM was analyzed by immunofluorescence staining of primary cultured granulosa cells (GCs). The body and ovarian weights decreased, the estrus cycles were erratic, and the FSH level increased, whereas the E2 level decreased in POF mice compared to controls. The pathological consequences of POF revealed an increase in atretic follicles, corpus luteum, and primordial follicles and a decrease in the number of primary, secondary, and tertiary follicles. Ki67 expression was reduced, β-gal, p21, p53, γH2AX, and pATM expression were elevated in the ovaries of POF mice. The expression of β-gal, γH2AX, and pATM increased in GCs with the concentration in a time-dependent manner. In total, CTX induced POF in mice, which was mediated by the DDR pathway of ATM-P53-P21.

The Effect of Prolactin on Gene Expression and the Secretion of Reproductive Hormones in Ewes during the Estrus Cycle.

Prolactin (PRL) plays an important role in animal follicle development and ovulation. However, its regulatory effects on the different stages of the estrus cycle in ewes are unclear. In this study, bromocriptine (BCR, PRL inhibitor) was used to study the effect of PRL on the secretion of reproductive hormones and gene expressions in order to explore its regulatory effects on the sexual cycle of ewes. Eighty healthy ewes with the same parity and similar weights were randomly assigned to the control group (C, n = 40) and the treatment group (T, n = 40, fed bromocriptine). After estrus synchronization, thirty-one ewes with overt signs of estrus were selected from each group. Six blood samples were randomly obtained by jugular venipuncture to measure the concentration of PRL, estrogen (E2), progesterone (P4), luteinizing hormone (LH), follicle-stimulating hormone (FSH), and gonadotropin-releasing hormone (GnRH) in the proestrus, estrus, metestrus, and diestrus. At the same time, we collected the ovaries of the six ewes in vivo after anesthesia in order to detect follicle and corpus luteum (CL) counts and measure the expression of hormone-receptor and apoptosis-related genes. The results show that PRL inhibition had no significant effects on the length of the estrus cycle (p > 0.05). In proestrus, the number of large and small follicles, the levels of E2, FSH, and GnRH, and the expressions of ER, FSHR, and the apoptotic gene Caspase-3 were increased (p < 0.05); and the number of middle follicles and the expression of anti-apoptotic gene Bcl-2 were decreased (p < 0.05) in the T group. In estrus, the number of large follicles, the levels of E2 and GnRH, and the expressions of the StAR, CYP19A1, and Bcl-2 genes were increased (p < 0.05), and the number of middle follicles was decreased (p < 0.05) in the T group. In metestrus, the number of small follicles and the expression of LHR (p < 0.05) and the pro-apoptotic gene Bax were increased (p < 0.05); the number of middle follicles was decreased (p < 0.05) in the T group. In diestrus, the number of large follicles, middle follicles, and CL, the level of P4, and the expressions of PR, 3β-HSD, StAR, Caspase-3, and Bax were increased (p < 0.05); the number of small follicles and the expression of Bcl-2 were decreased (p < 0.05) in the T group. In summary, PRL inhibition can affect the secretion of reproductive hormones, the follicle count, and the gene expression during the estrus cycle. These results provide a basis for understanding the mechanisms underlying the regulation of the ewe estrus cycle by PRL.

Administration of Nanocurcumin in Mice Models of Endometriosis as an Effort to Improve Folliculogenesis

The purpose of this study is to ascertain how nanocurcumin 0-50 nm affects malondialdehyde (MDA) levels, folliculogenesis, and the quantity of corpus luteum (CL) in a mice model of endometriosis. 24 mice were used in this scientific experiment, and they were split up into 4 treatment groups; positive control (C+) as mice model of endometriosis, Treatment 1 (T1) as mice model of endometriosis given nanocurcumin 0-50 nm (2.5 mg/KgBW) PO, Treatment 2 (T2) mice given nanocurcumin (5 mg/KgBW) and Treatment 3 (T3) as mice model of endometriosis treated with nanocurcumin (10 mg/KgBW). The nanocurcumin was administered for 14 days. The findings demonstrated that, out of all treatment groups, T3 had the lowest MDA level (20.78±4.58 ng/ml) and statistically significant (p&lt;0.05) compared to C+, T1 and T2. Significant differences (p&lt;0.05) were observed in the number of primary, secondary, and tertiary follicles at T3, from C+, T1, and T2, according to the folliculogenesis profile. Although the number of Graafian follicles tended to grow, there was no discernible difference between the number of Graafian follicles and the CL. It can be concluded that the administration of nanocurcumin for 14 days decreased MDA levels and increased the folliculogenesis profile. Administration of nanocurcumin at a dose of 10 mg/KgBW caused a decrease in MDA levels and an improvement in the folliculogenesis profile. Thus, administering nanocurcumin could improve the quality of folliculogenesis in endometriosis sufferers and improve reproductive health.

M6A demethylase FTO-stabilized exosomal circBRCA1 alleviates oxidative stress-induced granulosa cell damage via the miR-642a-5p/FOXO1 axis

BackgroundPremature ovarian insufficiency (POI) is an important cause of female infertility and seriously impacts the physical and psychological health of patients. Human umbilical cord mesenchymal stem cell-derived exosomes (HucMSCs-Exs, H-Exs) have exhibited protective effects on ovarian function with unclear mechanisms.MethodsA comprehensive analysis of the Gene Expression Omnibus (GEO) database were used to identify POI-associated circRNAs and miRNAs. The relationship between HucMSC-derived exosomal circBRCA1/miR-642a-5p/FOXO1 axis and POI was examined by RT-qPCR, Western blotting, reactive oxygen species (ROS) staining, senescence-associated β-gal (SA-β-gal) staining, JC-1 staining, TEM, oxygen consumption rate (OCR) measurements and ATP assay in vivo and in vitro. RT-qPCR detected the expression of circBRCA1 in GCs and serum of patients with normal ovarian reserve function (n = 50) and patients with POI (n = 50); then, the correlation of circBRCA1 with ovarian reserve function indexes was analyzed.ResultsHerein, we found that circBRCA1 was decreased in the serum and ovarian granulosa cells (GCs) of patients with POI and was associated with decreased ovarian reserve. H-Exs improved the disorder of the estrous cycles and reproductive hormone levels, reduced the number of atretic follicles, and alleviated the apoptosis and senescence of GCs in rats with POI. Moreover, H-Exs mitigated mitochondrial damage and reversed the reduced circBRCA1 expression induced by oxidative stress in GCs. Mechanistically, FTO served as an eraser to increase the stability and expression of circBRCA1 by mediating the m6A demethylation of circBRCA1, and exosomal circBRCA1 sponged miR-642a-5p to block its interaction with FOXO1. CircBRCA1 insufficiency aggravated mitochondrial dysfunction, mimicking FTO or FOXO1 depletion effects, which was counteracted by miR-642a-5p inhibition.ConclusionH-Exs secreted circBRCA1 regulated by m6A modification, directly sponged miR-642a-5p to upregulate FOXO1, resisted oxidative stress injuries in GCs and protected ovarian function in rats with POI. Exosomal circBRCA1 supplementation may be a general prospect for the prevention and treatment of POI.Graphical

Systemic low-dose anti-fibrotic treatment attenuates ovarian aging in the mouse.

The female reproductive system is one of the first to age in humans, resulting in infertility and endocrine disruptions. The aging ovary assumes a fibro-inflammatory milieu which negatively impacts gamete quantity and quality as well as ovulation. Here we tested whether the systemic delivery of anti-inflammatory (Etanercept) or anti-fibrotic (Pirfenidone) drugs attenuates ovarian aging in mice. We first evaluated the ability of these drugs to decrease the expression of fibro-inflammatory genes in primary ovarian stromal cells. Whereas Etanercept did not block Tnf expression in ovarian stromal cells, Pirfenidone significantly reduced Col1a1 expression. We then tested Pirfenidone in vivo where the drug was delivered systemically via mini-osmotic pumps for 6-weeks. Pirfenidone mitigated the age-dependent increase in ovarian fibrosis without impacting overall health parameters. Ovarian function was improved in Pirfenidone-treated mice as evidenced by increased follicle and corpora lutea number, AMH levels, and improved estrous cyclicity. Transcriptomic analysis revealed that Pirfenidone treatment resulted in an upregulation of reproductive function-related genes at 8.5 months and a downregulation of inflammatory genes at 12 months of age. These findings demonstrate that reducing the fibroinflammatory ovarian microenvironment improves ovarian function, thereby supporting modulating the ovarian environment as a therapeutic avenue to extend reproductive longevity.

The Effects of Saturated and Polyunsaturated Fatty Acids on Reproductive Performance and Reproductive Hormonal Changes in Dairy Cows at the Transition Period

Reproductive success is crucial in dairy farming as it heavily relies on the consumption of a complete mixed ration for the diet. The current study investigated the effects of adding saturated (SFA) and polyunsaturated fatty acids (PUFAs) to dairy cows' diets on reproductive performance and reproductive hormones during the transition period. A total of 30 Holstein dairy cows were randomly divided into three groups (10 animals in each group), based on parity and body condition score. The cows had an initial body weight of 567.5 ± 40.3 kg (mean ± SD), a body condition score of 3.5 ± 0.26 out of 5 (mean ± SD), and a parity of 1.7 ± 0.02 (mean ± SD). The control group received a balanced ration meeting all the nutrient requirements according to the National Research Council (NRC) guidelines. The SFA group received 1.4% of dry matter (DM) as palm oil (RumiFat®), while the omega group had 5% of DM as safflower (a source of n-6 fatty acids) added from 21 days before parturition to 21 days after, and 4% of DM as flaxseed (a source of n-3 fatty acids) added from 21 to 42 days after parturition. In the Omega group, estradiol concentration significantly increased on artificial insemination (AI) day (12.54 pg/mL). Additionally, serum prostaglandin F2-alpha concentration was significantly higher in the omega group (0.732 pg/mL on day 7 and 1.68 pg/mL on day 14) compared to other groups. The control group exhibited the highest progesterone concentrations at 14 and 21 days post-calving compared to the other groups, other groups. whereas the omega group highest concentration five days after AI. The omega group also showed a significantly higher mean number of follicles &gt;10mm and larger ovulatory follicle diameter. Moreover, a higher percentage of pregnant cows at 120 days in milk, fewer open days, and lower service per conception were observed in the omega group compared to the other groups. In conclusion, supplementing dairy cows' diets with PUFAs during the transition period positively influenced ovarian function, hormone levels, and reproductive performance.

Weightlessness leads to an increase granulosa cells in the growing follicle

The participation of women in space programs of increasing flight duration requires research of their reproductive system from the perspective of subsequent childbearing and healthy aging. For the first time, we present hormonal and structural data on the dynamics of recovery after a 157-day space flight in a woman of reproductive age. There were no clinically significant changes in the reproductive system, but detailed analysis shows that weightlessness leads to an increase in the proportion of early antral follicles and granulosa cells in large antral follicles. Returning to Earth’s gravity reduces the number and diameter of early antral follicles.

Molecular and transcriptomic analysis of the ovary during laying and brooding stages in Zhedong white geese (Anser cygnoides domesticus)

ABSTRACT 1. This study investigates the molecular mechanisms affecting brooding in Zhedong white geese by examining differences in reproductive endocrine levels, ovarian histology and transcriptomics. 2. Twenty 18-month-old Zhedong white geese were selected to examine their ovaries using histological, biochemical, molecular biological, and high-throughput sequencing techniques during the laying and brooding periods. 3. The results showed that the number of atretic follicles and apoptotic cells in the ovaries increased significantly (p < 0.05), the levels of follicle-stimulating hormone, luteinising hormone, gonadotropin-releasing hormone and oestradiol decreased significantly (p < 0.05), and the level of prolactin increased significantly (p < 0.01) during the brooding stage. 4. In broody geese, the expression of CASP3, CASP9, P53, BAX, and Cyt-c were considerably higher (p < 0.05), but BCL2 expression was significantly lower (p < 0.05). 5. In ovarian tissues, 260 differentially expressed lncRNAs, 13 differentially expressed miRNA and 60 differentially expressed mRNA were all discovered using transcriptome sequencing analysis. Functional enrichment analysis revealed that the differentially expressed mRNA and non-coding RNA target genes were primarily involved in ECM-receptor interaction, cell adhesion, cardiac muscle contraction, mTOR signalling, and the calcium signalling pathway. 6. In conclusion, follicular atrophy and apoptosis occurred in the ovaries and serum reproductive hormone levels were significantly changed during the brooding period of Zhedong white geese. COL3A1, COL1A2, GRIA1, RNF152, miR-192, and miR-194 may be important candidates for the regulation of brooding behaviour, with the mTOR signalling pathway playing a key role.

The effect of bisphenol A on the Notch (Notch2 and Jagged2) signaling pathway in the follicular development of the neonatal rat ovary

ABSTRACT The formation of primordial follicles determines the pool size of follicles in the ovary, and is crucial for female reproductivity. Oocyte nest breakdown, and the formation of primordial follicles, largely depend upon the communication between oocytes and the surrounding pregranulosa cells. The neurogenic locus notch homolog protein (Notch) signaling pathway is the key player for this cell-to-cell communication, and is responsible for primordial folliculogenesis. However, different endocrine disruptors, including bisphenol A (BPA; a plasticizer and a constituent of reusable plastic containers) may affect the Notch signaling pathway, and might induce ovary dysfunction via Notch signaling. Consequently, we investigated the possible influence of BPA treatment on the proportional distribution of the follicular stages, follicle numbers, levels of apoptosis, and on Notch2 and Jagged2 expressions in the ovary. BPA was administered at doses of either 50 µg/kg/day or 50 mg/kg/day, at different time intervals, during neonatal and fetal periods in vivo. After collecting the ovaries from the various experimental groups, follicles were counted, and frequency of apoptosis was determined by TUNEL assay. In addition, Notch2 and Jagged2 expressions were assessed by immunohistochemical staining and qPCR. In summary, BPA treatment affected the follicle numbers and apoptosis level, and Notch2 and Jagged2 expressions varied with follicular stage. It was also observed that these parameters were dose and time dependent with respect to BPA exposure.

Effect of serum anti-Müllerian hormone levels on the superovulation response in Holstein heifers.

Anti-Müllerian hormone (AMH) holds potential as a biomarker for assessing the superovulation (SO) response in cattle. Nonetheless, there exists scant information regarding this aspect in the literature concerning dairy heifers. Given this gap, our objective is to explore the viability of AMH as an indicator for gauging the SO response specifically in Holstein heifers. Furthermore, our aim encompasses examining the variations in AMH levels within the same individuals before and after undergoing SO. The study included 41 Holstein heifers. All heifers were superovulated and blood samples were taken both before and after the SO protocol. The findings revealed that the mean values of serum AMH concentrations before and after SO were 0.122ng/mL (0.093-0.248ng/mL) and 0.119ng/mL (0.084-0.170ng/mL), respectively. AMH concentrations in heifers were stratified into low (<0.106ng/mL), medium (0.107-0.126ng/mL) and high (>0.127ng/mL) categories both before and after SO. There was no significant correlation between AMH levels in the heifers both before and after SO treatment with the number of follicles, corpora lutea, total embryos collected or embryos transferred (p>0.05). Furthermore, this study showed that serum AMH concentrations in Holstein heifers did not change after SO treatment. In this study, as AMH levels in Holstein heifers were in a narrow range, a relationship between AMH and SO response could not be determined. In future studies, we believe that it would be more useful to plan more studies in Holstein donor heifers, taking into account the number of animals and AMH levels.

Structural gonadal lesions observed in Japanese quail (Coturnix coturnix japonica) following exposure during puberty to the neonicotinoid pesticide, imidacloprid

Exposure to the neonicotinoid insecticide, imidacloprid (IMI), causes reproductive toxicity in mammals and reptiles. However, reports on the effects of IMI on the gonads in birds are grossly lacking. Therefore, this study investigated the effects of pubertal exposure to IMI on the histology, ultrastructure, as well as the cytoskeletal proteins, desmin, smooth muscle actin and vimentin, of the gonads of Japanese quail (Coturnix coturnix japonica). Quails were randomly divided into four groups at 5 weeks of age. The control group was given only distilled water, whereas, the other three experimental groups, IMI was administered by oral gavage at 1.55, 3.1, and 6.2 mg/kg, twice per week for 4 weeks. Exposure to IMI doses of 3.1 and 6.2 mg/kg caused dose-dependent histopathological changes in the ovary and testis. In the ovary, accumulation of lymphocytes, degenerative changes, and necrosis with granulocyte infiltrations were observed, while in the testis, distorted seminiferous tubules, germ cell sloughing, vacuolisations, apoptotic bodies, autophagosomes, and mitochondrial damage were detected. These changes were accompanied by a decreased number of primary follicles (P ≤ 0.05) in the ovary and a decrease (P ≤ 0.05) in the epithelial height, luminal, and tubular diameters of seminiferous tubules at the two higher dosages. In addition, IMI had a negative effect on the immunostaining intensity of desmin, smooth muscle actin, and vimentin in the ovarian and testicular tissue. In conclusion, exposure to IMI during puberty can lead to a range of histopathological alterations in the gonads of Japanese quails, which may ultimately result in infertility.

Atorvastatin improves ovarian function and follicular reserve in rats with premature ovarian insufficiency

Research questionCan atorvastatin with antioxidant, anti-inflammatory, and anti-apoptotic properties improve ovarian function and follicular reserve in a model of premature ovarian insufficiency induced by cyclophosphamide? DesignIn this experimental study, twenty-four adult female Wistar rats were divided into four groups: control, POI, POI + atorvastatin, and atorvastatin. After treatment with atorvastatin, the serum levels of glutathione, total antioxidant capacity, malondialdehyde, follicle-stimulating hormone, estradiol, anti-Müllerian hormone, tumor necrosis factor-alpha, and interleukin-6 were evaluated. Additionally, the mRNA and protein expression of Bax, Bcl-2, and VEGF-A, the number of follicles, and the volume of the ovary were examined. ResultsOur result showed that the serum levels of total antioxidant capacity (P < 0.001), glutathione, estradiol,and anti-Müllerian hormone (P < 0.05), the expression of Bcl-2 and VEGF-A (P < 0.05), the number of primordial and primary follicles (P < 0.001), preantral and antral follicles (P < 0.01), the total volume of the ovary, and the volume of the cortex (P < 0.05) in the POI + atorvastatin group compared to the POI group increased significantly, while the serum levels of malondialdehyde, tumor necrosis factor-alpha, and interleukin-6, and the expression of Bax were significantly decreased (P < 0.05) ConclusionsAtorvastatin can significantly reduce the detrimental effects of cyclophosphamide in the POI model by reducing oxidative stress and proinflammatory cytokines, as well as regulating the expression of Bax, Bcl-2, and VEGF-A and improves ovarian function and follicular reservation.